Pharmaceutical composition containing berberine as effective ingredient for preventing and treating addiction or tolerance to morphine

ABSTRACT

Disclosed is a pharmaceutical composition for preventing and treating addiction to morphine or preventing and inhibiting the development of tolerance to the analgesic effects of morphine, containing berberine as an effective ingredient, wherein the berberine has an inhibitory effect versus psychological dependence on abused drugs such as morphine and the increase of spontaneous locomotor activity upon administration of the drugs, The pharmaceutical composition and a  Coptis japonica  plant extract of the present invention, which contain berberine, are highly effective in inhibiting the aforementioned symptoms of morphine addiction, and are thus useful for prevention and treatment of addiction to abused drugs such as morphine. In addition, the pharmaceutical composition additionally containing a pharmaceutical acceptable carrier can be applied to prevent and inhibit morphine tolerance caused by repeated administration of morphine, while not affecting the analgesic effects of morphine upon a single administration.

TECHNICAL FIELD

The present invention, in general, relates to a pharmaceuticalcomposition for preventing and treating the harmful actions of narcoticanalgesic agents. More particularly, the present invention relates to apharmaceutical composition comprising berberine as an effectiveingredient, where the composition has inhibitory effects versuspsychological dependence on morphine and an increase of spontaneouslocomotor activity induced by morphine.

People who abuse drugs typically suffers from psychological dependencewhich is a compulsion for the continuous administration of a drug forits euphoric effects despite any adverse effects that may occur, andinclude morphine or derivatives thereof, cocaine, and methamphetamine orderivatives thereof.

Morphine, which is a narcotic analgesic agent, has the psychologicalexcitatory effects with most common symptoms of hallucinosis anddelirium when used in relatively high doses or when repeatedly used forlonger periods even in low doses. Such psychotoxicity occurrence isaugmented by the repeated administration of morphine, and a very highphysical and psychological dependence is thus developed. That is, theabused drugs have the major properties of exciting the central nervoussystem and increasing the psychological craving for continuousconsumption of the drugs. The abused drugs increase spontaneouslocomotor activity by continuous administration and induce psychologicaldependence thereon.

The continuous administration of the abused drugs leads to theexhaustion of dopamine and reduced activity of dopamine in the nervoussystem. To compensate for the reduced activity of dopamine, dopamineneurons are activated. Thus, the postsynaptic dopamine receptors becomehypersensitive, and spontaneous locomotor activity is increased,resulting in the development of strong craving for the continuousconsumption of the drugs. Due to such a psychoactive effect of morphine,users depending on morphine have been continuously increased. Since suchabuse of morphine causes severe social problems, there is an urgent needfor the development of agents for treating and preventing morphineaddiction.

PRIOR ART

Korean Pat. No. 10-0277481 discloses isatinoxime derivatives, whichserve as antagonists versus neurotoxic effects of excitatory amino acidsand thus are useful for treatment of excitatory amino acid-dependentdiseases in the central nervous system, cerebrovascular diseases, mentalillness, etc. However, currently, there is no development for drugs withobvious therapeutic and preventory efficacies on an increase ofspontaneous locomotor activity and psychological dependence, which arecaused by the abused drugs such as morphine. Thus, the abused drugs areproblematic in terms of having harmful side effects such as the increaseof spontaneous locomotor activity and the development of psychologicaldependence. In particular, the abuse and addiction of the drugs have ledto severe social problems.

On the other hand, analgesic agents are medical drugs that are typicallyused for inhibiting or alleviating acute pains. The analgesic agents aretypically classified into narcotic and non-narcotic types. The narcoticanalgesic agents have been clinically used for treating acute pains inlate-stage cancer patients due to their excellent analgesic effectversus visceral pains. However, as described above, when the narcoticanalgesic agents are administered in relatively high doses or repeatedlyadministered for longer periods even in low doses, the habitualadministration of the analgesics and the craving for the analgesicagents can be developed. Also, when a user is repeatedly exposed to anarcotic analgesic agent of a predetermined amount, tolerance to theanalgesic agent is rapidly developed, resulting in the reduction of theanalgesic effect of the analgesic. Thus, the narcotic analgesic agentshould be used in an increased dose in order to obtain a desiredanalgesic effect. Such an increased dose of the narcotic analgesic agentcauses different side effects and, thus, is severely toxic to humans.

As a representative example of the narcotic analgesic agent, morphine isknow to have the highest analgesic effect among analgesics developed todate. However, the repeated administration of morphine causes gradualreduction in its analgesic effect and finally the development oftolerance to the analgesic effect of morphine. For these reasons,morphine is limited in its application to the clinical fields despiteits excellent efficacy. Therefore, in order to utilize morphine as ananalgesic agent, it is very important that the morphine's initialanalgesic effect is maintained without reduction in the analgesic effectupon repeated administration. This purpose can be achieved by inhibitingthe development of tolerance to morphine.

A pharmaceutical composition for preventing or inhibiting thedevelopment of tolerance to the analgesic effect of the narcoticanalgesics such as morphine is described in Korean Pat. Publication No.1999-0036248 (Application No. 10-1998-0700916), which discloses apharmaceutical composition with an inhibitory effect versus thedevelopment of psychological dependence and/or tolerance for narcoticanalgesics, comprising 2-(1-pyrrolidinyl)acetateamide derivatives aseffective ingredients. However, with respect to components ofcompositions, the above pharmaceutical composition differs from thepresent pharmaceutical composition comprising berberine. Also, apharmaceutical composition containing berberine is disclosed in KoreanPat. No. 10-0281003, which, in detail, comprises protoberberine alkaloidcompounds as effective ingredients and has an anti-depressing effect.However, this composition that may be used as anti-depressant differsfrom the present pharmaceutical composition having an inhibitory effectversus morphine tolerance by the action of berberine.

DISCLOSURE OF THE INVENTION

Leading to the present invention, the intensive and thorough researchonto to develop preventory or therapeutic agents for addiction to theabused drugs, conducted by the present inventors, resulted in thefinding that naturally occurring berberine, a major component of Coptisjaponica and Phellodendron amurense plants, has a significant inhibitoryeffect versus psychological dependence in an animal model with inducedpsychological dependence.

It is therefore an object of the present invention to provide apharmaceutical composition for preventing and treating morphineaddiction.

Also, the present inventors attempted to develop pharmaceuticalpreparations having an inhibitory effect versus the morphine tolerancecaused by repeated administration of morphine. This research resulted inthe finding that naturally occurring berberine, a major component ofCoptis japonica, Phellodendron amurense and Berberis koreana plants, hasa strong inhibitory effect versus morphine tolerance.

Therefore, it is another object of the present invention to provide apharmaceutical composition for preventing and inhibiting the developmentof morphine tolerance, comprising berberine as an effective ingredient.

BRIEF DESCRIPTION OF THE DRAWINGS

The above and other objects, features and other advantages of thepresent invention will be more clearly understood from the followingdetailed description taken in conjunction with the accompanyingdrawings, in which:

FIG. 1 is a graph showing an inhibitory effect of berberine versuspsychological dependence caused by berberine in mice;

FIG. 2 is a graph showing an inhibitory effect of berberine versusspontaneous locomotor activity increased by berberine in mice;

FIG. 3 is a graph showing an effect of berberine on the analgesic effectof morphine when morphine was administered once to mice; and

FIG. 4 is a graph showing an inhibitory effect of berberine versustolerance to the analgesic effect of morphine when morphine wasrepeatedly administered to mice.

BEST MODES FOR CARRYING OUT THE INVENTION

The meaning of the abbreviations on the drawings are as follows:

sec = second SAL = saline BER = berberine MOR = morphine AUC = areaunder the curve DW = distilled water.

The present invention provides a novel use of berberine and apharmaceutical composition comprising berberine. More particularly, thepresent invention provides a pharmaceutical composition for preventingand treating morphine addiction or preventing and inhibiting thedevelopment of tolerance to the analgesic effect of morphine, comprisingberberine as an effective ingredient.

In an aspect of the present invention, there is provided apharmaceutical composition for preventing and inhibiting morphineaddiction and the development of tolerance to the analgesic effect ofmorphine, comprising a pharmaceutically acceptable carrier along withthe above effective ingredient.

In another aspect of the present invention, there is provided a use ofberberine for preparation of a pharmaceutical composition for preventingand inhibiting morphine addiction and the development of tolerance tothe analgesic effect of morphine.

In the pharmaceutical composition of the present invention, theeffective ingredient berberine(7,8,13,13a-tetradihydro-9,10-dimethoxy-2,3-(methyldioxy)berbinium),belonging to the isoquinolineakaloid family, can be extracted from avariety of plants, for example, from the root of Coptis japonica and thebark of Phellodendron amurense, where the two plants belong to thefamily Berberidaceae. The berberine was used for staining wool, silk,leather, and the like. The berberine, currently used for medicalpurposes, is known to have antibacterial, intestine-regulating andanti-ulcer effects. Safe doses of the berberine are described in theliterature, and the berberine is known to have a LD50 value of 90 mg/kg(intraperitoneal injection) in rats (see, Tang, W. and Eisenbrand, G.,Chinese Drugs of Plant Origin, pp. 362-371, Springer Verlag, Berlin,Heidelberg, N.Y.). The berberine used in the pharmaceutical compositionof the present invention is commercially available.

The daily dosage of the berberine used as an effective ingredient in thepresent pharmaceutical composition may be determined depending onpatient's age, body weight and pathological states, and typically rangesfrom 20 to 100 mg (intraperitoneal injection) or 50 to 400 mg (oraladministration). Also, the dosage of the berberine may be properlydetermined by experience of those skilled in the art. The berberine iscurrently used as a medical drug, and known to rarely have toxicity whenadministered to the body in the typical doses. Therefore, it will beunderstood by those of ordinary skills in the art that thepharmaceutical composition for preventing and inhibiting morphineaddiction and the development of tolerance to the analgesic effect ofmorphine, comprising an effective amount of berberine, is safe becauseof not having harmful side effects to the humans.

In the present invention, a narcotic analgesic agent is exemplified byonly morphine in the above part of this specification. However, iftolerance to the narcotic analgesic agent is induced by singleadministration or repeated administration for short or long periods, thenarcotic analgesic agent is not limited to morphine. Non-limitingexamples of the narcotic analgesic include morphine derived from opiumand its semi-synthetic derivatives, and non-natural compounds having themorphine-like effect, such as pethidine, and salts thereof. In moredetail, the narcotic analgesic agent is exemplified by alkaloids derivedfrom opium and their semi-synthetic derivatives such as phenanthrenes(e.g., morphine, oxymorphone, hydromorphone, codeine, hydrocodone,heroin, thebaine and buprenorphine); phenylpiperidines (e.g., meperidineand fentanyl); phenylheptylamine (e.g., methadone and propoxylphene);morphinans (e.g.; levolphanol, methorphan and levolphan); andbenzomorphans (e.g., phenazocine and pentazocine).

The present pharmaceutical composition comprising berberine, accordingto its formulation, may further comprise a pharmaceutically acceptablecarrier commonly used in the art. In detail, the present pharmaceuticalcomposition comprising berberine may be administered in the form of oralpreparations or injection preparations. Examples of the oralpreparations may include tablets and gelatin capsules. The oralpreparations, in addition to the active ingredient, may comprise adiluent (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, celluloseand/or glycine), and a lubricant (e.g., silica, talc, stearic acid andmagnesium or calcium salts thereof and/or polyethylene glycol).Preferably, the tablets may further comprise a binder (e.g., magnesiumaluminum silicate, starch paste, gelatin, tragacanth, methylcellulose,sodium carboxymethylcellulose and/or polyvinylpyrolidine), and, ifdesired, may further comprise a disintegrator (e.g., starch, agar,arginic acid or sodium salts thereof, or a mixture thereof), and/or anabsorbent, a colorant, a flavoring agent and a sweetening agent. Theinjection preparations may be preferably an isotonic solution or asuspension, and may be sterilized or comprise an adjuvant (e.g., anantiseptic, a stabilizer, a wetting agent or an emulsifier, a salt forosmotic regulation and/or a buffering agent). In addition, theformulations may further comprise other therapeutically usefulsubstances.

The berberine as an effective ingredient of the pharmaceuticalcomposition of the present invention was found not to affect theanalgesic effect of morphine upon a single administration. In addition,the berberine was found to have an effect of significantly inhibitingthe development of tolerance to the analgesic effect of morphine inducedby repeated administration while maintaining the initial analgesiceffect of morphine. Therefore, the pharmaceutical composition of thepresent invention may be used to prevent the development of tolerance tothe analgesic effect of morphine upon administration with morphine.Consequently, the present pharmaceutical composition may be used with apurpose for preventing the development of morphine tolerance. Inaddition, the present pharmaceutical composition has an effect ofinhibiting and alleviating tolerance to the analgesic effect ofmorphine, induced by repeated administration with morphine. Thus, thepresent pharmaceutical composition may be therapeutically used with anaim to inhibit the previously induced tolerance to morphine, along withmorphine administration.

The present invention will be explained in more detail with reference tothe following examples in conjunction with the accompanying drawings.However, the following examples are provided only to illustrate thepresent invention, and the present invention is not limited to theexamples.

EXAMPLES Method and Materials

Male ICR mice weighing 18 to 25 g were adapted to a new environment bybeing grown for over one week in a breeding room under a controlledtemperature and humidity. 10 to 15 mice per group were used. Morphineand berberine were purchased from the Keukdong Pharm Company (Inchon,Korea) and the Sigma Company (USA), respectively.

Example 1 Evaluation of Inhibitory Effect of Berberine Versus theDevelopment of Psychological Dependence for Morphine

The development of psychological dependence on morphine was detected bythe conditioned place preference test.

Conditioned place preference was performed in two boxes of equal size(15×15×15 cm) were used, each which has a forehead surface made of atransparent acrylic plate. The remaining three surfaces of each box weremade of a white or black acrylic plate. The two boxes were connectedwith a gray passage (3×3×7.5 cm), and the passage could be blocked witha removable gillotine door. To allow the mice to feel the texture of thefloor of the boxes, the white box had a tough floor, and the black boxhad a smooth floor. The mice were grown under a light intensity of 20Lux.

Step 1 (Preconditioning Phase)

On day 1, the guillotine door of the boxes was open, and the mice wereallowed to explore both compartments freely for 5 min. On day 2, themice were placed in the boxes in the same way as in day 1, and the timesspent in both boxes were measured for 15 min and used as controllatency.

Step 2 (Conditioning Phase)

On days 3, 5, 7 and 9, the guillotine door was closed. The mice wereintraperitoneally administered with 5 mg/kg of morphine, and placed forone hour in the white box with the hatred effect. On days 4, 6, 8 and10, the mice were administered with physiological saline and placed forone hour in the black box with the preference effect. One hour beforethe morphine administration, 1 or 2 mg/kg of berberine was orallyadministered to the mice.

Step 3 (Testing Phase)

On day 9, after the guillotine door was opened, the times the mice spentin the boxes were measured for 15 min, wherein the mice were notadministered with any drug, and compared to those measured on day 2. Thedegree of the development of psychological dependence was calculated bysubtracting the preconditioning time from the testing time. As shown inFIG. 1, a morphine control showed a significant psychological dependence(p<0.01). In contrast, in the mice administered with morphine andberberine, the psychological dependence was completely inhibited(p<0.01) to the level equal to the control treated with physiologicalsaline. These results indicate that berberine has an excellentinhibitory effect versus psychological dependence on morphine.

Example 2 Evaluation of Inhibitory Effect of Berberine Versus Increaseof Spontaneous Locomotor Activity by Morphine

Spontaneous locomotor activity was tested in a plastic chamber (26×30×30cm) using a Video-tracking system. First, the mice were administeredwith 10 mg/kg of morphine once per day for six days. Immediately afterthe last administration, the mice were placed in the plastic chamber,and spontaneous locomotor activity was recorded for 30 min and analyzedusing a computer program. Berberine was orally administered to the micein amounts of 1 and 2 mg/kg one, hour before administration of morphine.The results are given in FIG. 2. As shown in FIG. 2, when administeredwith only morphine, the mice showed a remarkably increased locomotoractivity of 50242 steps by a forefoot. In contrast, whenpre-administered with 1 and 2 mg/kg of berberine, the mice displayedlocomotor activities of 19744 steps and 20027 steps. That is, berberinewas found to significantly suppress the locomotor activity increased bymorphine by 61% and 60% (p<0.001, respectively) in comparison with themorphine control group.

Example 3 Evaluation of the Effects of Berberine on the Analgesic Effectof Morphine Upon a Single Administration

Male ICR mice weighing 18 to 25 g were adapted to a new environment bybeing grown for over one week in a breeding room under controlledtemperature and humidity. 10 mice per group were used. Morphine andberberine (berberine hemisulfate) were purchased from the Keukdong PharmCompany (Inchon, Korea) and the Sigma Company (USA), respectively, anddissolved in distilled water before use.

The effect of berberine on the analgesic action of morphine was assessedby a hot-plate test applying a heat stimulus to mice. For the hot-platetest, each mouse was placed on a hot plate at 52° C. and the latencyuntil mouse showed first signs of discomfort (hind paw-licking orjumping) was observed. To avoid tissue damage, an artificial maximumtime for exposure to heat was imposed (cut-off time), which was 30 sec.In case of not showing the behavioral response for over 30 sec, micewere taken off the hot plate. The mice were intraperitoneallyadministered with distilled water and berberine of 1, 3 and 10 mg/kg.After 30 min, the mice were subcutaneously injected with distilled waterand morphine of 5 mg/kg. 30, 60 and 90 min after the injection, lickingor jumping responses of the mice to heat from the hot plate was measuredaccording to the same method as in the recording of the latency, with amaximum time of 30 sec. The results are expressed as “percentage maximalpossible effect (% MPE)”, which was calculated according to an equation,below, and the strength of the analgesic action of morphine wasexpressed as “Area Under the Curve (AUC)”.MPE (%)=(Tt−To)/(Tc−To)×100

wherein, Tc is the cut-off time, Tt is the test latency, and To is thecontrol latency.

As shown in FIG. 3 (significant difference between control andmorphine-treated group: ***P<0.001), the morphine-treated group showed asignificantly increased analgesic effect, compared to the control grouptreated with physiological saline. In addition, in the groupspre-treated with berberine of 1, 3 and 10 mg/kg, berberine was found torarely affect the analgesic effect of morphine.

Example 4 Evaluation of Effects of Berberine Versus the Development ofMorphine Tolerance Caused by Repeated Administration

Berberine was evaluated for an effect on the development of morphinetolerance by repeated administration. Mice were pretreated withdistilled water and berberine of 1, 3 and 10 mg/kg by intraperitonealinjections. After 30 min, the mice were subcutaneously injected withphysiological saline and morphine of 10 mg/kg once per day for six days.The next day, that is, on day 7, behavioral responses of the mice toheat on a hot plate were observed according to the same hot-plate testas in the Example 3. The development of tolerance to the analgesiceffect of morphine was expressed as AUC, which was calculated accordingto the same method as in the Example 3.

As shown in FIG. 4 (significant difference between control andmorphine-treated group with a single administration: ***P<0.01;significant difference between control and morphine-treated group withmorphine tolerance: ⁺P<0.05; significant difference between group withmorphine tolerance and berberine-treated group: ⁺⁺P<0.01), when micewere repeatedly administered with morphine, the initial analgesic effectof morphine was greatly reduced, indicating that morphine tolerance wasdeveloped. In contrast, in the group pretreated with berberine of 10mg/kg, such morphine tolerance was found to be significantly inhibited.

INDUSTRIAL APPLICABILITY

As described hereinbefore, berberine nearly completely recovered themorphine-poisoned mice to the level of normal mice (physiologicalsaline-treated group), indicating that berberine has an effect ofgreatly alleviating addiction to morphine. In addition, berberinesignificantly inhibited the increase of spontaneous locomotor activityby repeated administrations of morphine. Therefore, berberine is usefulfor the prevention and treatment of morphine addiction.

In addition, the pharmaceutical composition of the present invention hasthe excellent effects on prevention and inhibition of the development oftolerance to the analgesic effect of morphine upon repeatedadministration, while not affecting the analgesic effect of morphineupon a single administration. Therefore, the present composition is veryuseful for preventing or alleviating the development of tolerance tomorphine.

1. A method for inhibiting a person from development of tolerance toanalgesic effects of morphine comprising administering to such person apharmaceutical composition having berberine as an effective ingredient.2. A method according to claim 1, wherein the pharmaceutical compositioncomprises a pharmaceutically acceptable carrier.